安全与环境学报
安全與環境學報
안전여배경학보
JOURNAL OF SAFETY AND ENVIRONMENT
2010年
1期
98-101
,共4页
杨健%赵化冰%张明露%朱滨%朱琳%蔡宝立
楊健%趙化冰%張明露%硃濱%硃琳%蔡寶立
양건%조화빙%장명로%주빈%주림%채보립
微生物学%海水%创伤弧菌%PCR%实时定量PCR%ITS序列
微生物學%海水%創傷弧菌%PCR%實時定量PCR%ITS序列
미생물학%해수%창상호균%PCR%실시정량PCR%ITS서렬
microbiology%seawater%Vibrio vulnificus%PCR%real-time PCR%ITS sequence
采用普通PCR方法和SYBR Green实时定量PCR方法,根据创伤弧菌16S rDNA和23S rDNA基因间隔序列(internal transcribed spacer,ITS)设计引物,对渤海湾天津沿岸海水中的创伤弧菌进行了定性和定量检测.8个样品采集自天津市汉沽区沿岸海水,采集时间分别为2008年7月、10月和2009年3月、5月.PCR检测结果表明,这些样品都能扩增出276 bp的ITS序列,这些序列和GenBank上的同源序列(DQ462478)的相似性为96%~98%.用SYBR Green实时定量PCR方法测定了8个海水样品,样品中创伤弧菌的浓度为7.12×10~3~8.40×10~5 个/L,表明天津沿岸海水存在严重的创伤弧菌污染.
採用普通PCR方法和SYBR Green實時定量PCR方法,根據創傷弧菌16S rDNA和23S rDNA基因間隔序列(internal transcribed spacer,ITS)設計引物,對渤海灣天津沿岸海水中的創傷弧菌進行瞭定性和定量檢測.8箇樣品採集自天津市漢沽區沿岸海水,採集時間分彆為2008年7月、10月和2009年3月、5月.PCR檢測結果錶明,這些樣品都能擴增齣276 bp的ITS序列,這些序列和GenBank上的同源序列(DQ462478)的相似性為96%~98%.用SYBR Green實時定量PCR方法測定瞭8箇海水樣品,樣品中創傷弧菌的濃度為7.12×10~3~8.40×10~5 箇/L,錶明天津沿岸海水存在嚴重的創傷弧菌汙染.
채용보통PCR방법화SYBR Green실시정량PCR방법,근거창상호균16S rDNA화23S rDNA기인간격서렬(internal transcribed spacer,ITS)설계인물,대발해만천진연안해수중적창상호균진행료정성화정량검측.8개양품채집자천진시한고구연안해수,채집시간분별위2008년7월、10월화2009년3월、5월.PCR검측결과표명,저사양품도능확증출276 bp적ITS서렬,저사서렬화GenBank상적동원서렬(DQ462478)적상사성위96%~98%.용SYBR Green실시정량PCR방법측정료8개해수양품,양품중창상호균적농도위7.12×10~3~8.40×10~5 개/L,표명천진연안해수존재엄중적창상호균오염.
The paper is aimed at introducing our research on the way of how to detect and quantify Vibrio vulnificus in Tianjin near-coast seawater of Bohai Bay by means of conventional PCR and SYBR green real-time quantitative PCR with the primer pair based on 16S rDNA-23S rDNA internal transcribed spacer (ITS) of Vibrio vulnificus. For our research purpose, we have chosen eight samples from Hangu, one of the above said seawater areas in Tianjin, in July and Oct., 2008 and Mar. And May, 2009. PCR analysis shows that a 276 bp ITS sequence was amplified from all the eight seawater samples and the sequential results show that identity of these DNAs to the homologous sequences in the GenBank (DQ462478) were 96%-98%. In addition, the SYBR Green real-time quantitative PCR analysis also indicates that Vibrio vulnificus concentration in the above-mentioned samples range from 7.12×10~3 cells/L to 8.40×10~5 cells/L, suggesting that there is a serious water contamination problem in Tianjin coastal seawater. The entire work of detection and qualitative analysis of Vibrio vulnificus, including sample processing, extraction of bacterial DNA, conventional and real-time PCR amplifications, proves able to get finished in 12 hours, making it a rapid single-day assay.